Occurrence, molecular characterization and antibiogram of water quality indicator bacteria in river water serving a water treatment plant

Water pollution by microorganisms of fecal origin is a current world-wide public health concern. Total coliforms, fecal coliforms (Escherichia coli) and enterococci are indicators commonly used to assess the microbiological safety of water resources. In this study, influent water samples and treated water were collected seasonally from a water treatment plant and two major water wells in a Black Belt county of Alabama and evaluated for water quality indicator bacteria. Influent river water samples serving the treatment plant were positive for total coliforms, fecal coliforms (E. coli), and enterococci. The highest number of total coliform most probable number (MPN) was observed in the winter (847.5 MPN/100 mL) and the lowest number in the summer (385.6 MPN/100 mL). Similarly E. coli MPN was substantially higher in the winter (62.25 MPN/100 mL). Seasonal variation of E. coli MPN in influent river water samples was strongly correlated with color (R² = 0.998) and turbidity (R² = 0.992). Neither E. coli nor other coliform type bacteria were detected in effluent potable water from the treatment plant. The MPN of enterococci was the highest in the fall and the lowest in the winter. Approximately 99.7 and 51.5 enterococci MPN/100 mL were recorded in fall and winter seasons respectively. One-way ANOVA tests revealed significant differences in seasonal variation of total coliforms (P < 0.05), fecal coliforms (P < 0.01) and enterococci (P < 0.01). Treated effluent river water samples and well water samples revealed no enterococci contamination. Representative coliform bacteria selected by differential screening on Coliscan Easygel were identified by 16S ribosomal RNA gene sequence analysis. E. coli isolates were sensitive to gentamicin, trimethoprim/sulfamethazole, ciprofloxacin, vancomycin, tetracycline, ampicillin, cefixime, and nitrofurantoin. Nonetheless, isolate BO-54 displayed decreased sensitivity compared to other E. coli isolates. Antibiotic sensitivity pattern can be employed in microbial source tracking.     

Pilot-plant comparative study of peracetic acid and sodium hypochlorite wastewater disinfection

Peracetic acid (PAA) use in wastewater disinfection was assessed by examining its performances in a pilot plant fed by the effluent from a conventional activated-sludge treatment plant. The influence of PAA initial concentrations (0.5-4.0 mg/l) and contact times (8-38 min) on the presence of seven microorganisms (total coliforms, fecal coliforms, fecal streptococci, Escherichia coli, Pseudomonas sp., Salmonella sp., and bacteriophages anti-E. coli) and on residual biocide and halogenated organic compound (AOXs) concentrations were evaluated. The data so obtained were compared to the corresponding results acquired using sodium hypochlorite (HYP) in the same experimental conditions. The biocide effect of PAA against total and fecal coliforms, E. coli, Pseudomonas sp. and Salmonella sp. was similar to that shown by HYP. The former disinfectant was, however, less efficient than the latter in the reduction of fecal streptococci and bacteriophages anti-E. coli. In both cases the biocide quantities initially introduced in the sewage resulted in the presence of significant concentrations at the end of the contact time. No significant variation of AOX content was detected in the effluent treated with PAA, whereas a progressive increment of such compounds was found when increasing quantities of HYP were added to the sewage.     

The limitation of the ratio of fecal coliforms to total coliphage as a water pollution index

The fecal coliform populations of raw sewage, sewage lagoon effluent, and river water were determined using the most probable number technique. The total coliphage populations of the three water sources were determined using Escherichia coli B (ATCC 11303-1) host cells. The ratios of fecal coliforms to coliphage in the three water samples were 87:1, 4.2:1, and 0.15:1, respectively. The ratio of fecal coliforms to coliphage in stored raw sewage decreased from 87:1 to about 1:1 within 7 days at 20°C and within 28 days at 4°C. These changing ratios resulted from the greater longevity of the coliphage compared with that of the coliform bacteria. The use of the ratio of fecal coliforms to coliphage is not considered reliable as an index of when a fecal pollution event occurred because the ratio is influenced by prior contamination, presence of sediment, chlorination, and temperature.     

Confirmation of E. coli among other thermotolerant coliform bacteria in paper mill effluents, wood chips screening rejects and paper sludges

Paper sludges are solid wastes material generated from the paper production, which have been characterized for their chemical contents. Some are rich in wood fiber and are a good carbon source, for example the primary and de-inking paper sludges. Others are made rich in nitrogen and phosphorus by pressing the activated sludge, resulting from the biological water treatments, with the primary sludge, yielding the combined paper sludge. Still, in the absence of sanitary effluents very few studies have addressed the characterization of their coliform microflora. Therefore, this study investigated the thermotolerant coliform population of one paper mill effluent and two paper mill sludges and wood chips screening rejects using chromogenic media. For the first series of analyses, the medium used was Colilert broth and positive tubes were selected to isolate bacteria in pure culture on MacConkey agar. In a second series of analyses, double selective media, based on ss-galactosidase and ss-glucuronidase activities, were used to isolate bacteria. First, the presence of thermotolerant coliforms was detected in low numbers in most water effluents, but showed that the entrance of the thermotolerant coliforms was early in the industrial process. Also, large numbers of thermotolerant coliforms, i.e., 7,000,000 MPN/g sludge (dry weight; d.w.), were found in combined sludges. From this first series of isolations, bacteria were purified on MacConkey medium and identified as Citrobacter freundii, Enterobacter sp, E. sakazakii, E. cloacae, Escherichia coli, K. pneumoniae, K. pneumoniae subsp. rhinoscleromatis, K. pneumoniae subsp. ozaenae, K. pneumoniae subsp. pneumoniae, Pantoea sp, Raoultella terrigena, R. planticola. Second, the presence of thermotolerant coliforms was measured at more than 3,700-6,000 MPN/g (d.w) sludge, whereas E. coli was detected from 730 to more than 3,300 MPN/g (d.w.) sludge. The presence of thermotolerant coliform bacteria and E. coli was sometimes detected from wood chips screening rejects in large quantities. Also, indigenous E. coli were able to multiply into the combined sludge, and inoculated E. coli isolates were often able to multiply in wood chips and combined sludge media. In this second series of isolations, API20E and Biolog identified most isolates as E. coli, but others remained unidentified. The sequences of the 16S rDNA confirmed that most isolates were likely E. coli, few Burkholderia spp, but 10% of the isolates remained unidentified. This study points out that the coliform bacteria are introduced by the wood chips in the water effluents, where they can survive throught the primary clarifier and regrow in combined sludges.     

Distribution of indicator bacteria in Canyon Lake, California

The spatial and temporal distributions of indicator bacteria in a small, multiple-use source drinking water reservoir in Southern California, USA were quantified over the period August 2001-July 2002. High levels of total and fecal coliform bacteria were present in Canyon Lake (annual geometric mean concentrations+/-SEM of 3.93+/-0.02 and 3.02+/-0.03 log cfu/100mL, respectively), while comparatively low levels of enterococci and E. coli were found (1.16+/-0.02 log cfu/100mL and 0.30+/-0.03 log MPN/100mL, respectively). As a result, these different indicator bacteria yielded quite divergent indices of water quality, with 72.1% of all surface samples (n=294) exceeding the USEPA single-sample limit of 400 cfu/100mL fecal coliform bacteria, while none (0%) of the samples exceeded the single-sample limit for E. coli (n=194). Regression analyses found a positive correlation between total and fecal coliform bacteria (R=0.50, significant at p<0.001) and between enterococci and E. coli (R=0.51, significant at p<0.001), but no correlation or inverse correlations were found between coliform concentrations and enterococci and E. coli levels. External sources responsible for the high total and fecal coliform bacteria were not identified, although laboratory studies demonstrated growth of the coliform bacteria in lake water samples. Enterococci and E. coli were not observed to grow, however. Bacteria concentrations varied relatively little laterally across the lake, although strong vertical gradients in fecal coliform and enterococcus bacteria concentrations were present during summer stratification, with concentrations about 10x higher above the thermocline when compared with surface concentrations. In contrast, total bacteria, total virus and total coliform bacteria levels were unchanged with depth. Seasonal trends in bacteria concentrations were also present. This study shows that the choice of indicator bacteria and sampling depth can both strongly affect the apparent microbial water quality of a lake or reservoir.     

Disinfection efficiency of peracetic acid, UV and ozone after enhanced primary treatment of municipal wastewater

The City of Montreal Wastewater Treatment Plant uses enhanced physicochemical processes (ferric and/or alum coagulation) for suspended solids and phosphorus removal. The objective of this study was to assess the ability of peracetic acid (PAA), UV, or ozone to inactivate the indicator organisms fecal coliforms, Enterococci, MS-2 coliphage, or Clostridium perfringens in the effluent from this plant. PAA doses to reach the target fecal coliform level of 9000 CFU/100mL exceeded 6 mg/L; similar results were obtained for enterococci, and no inactivation of Clostridium perfringens was observed. However a 1-log reduction of MS-2 occurred at PAA doses of 1.5 mg/L and higher. It was expected that this effluent would have a high ozone demand, and would require relatively high UV fluences, because of relatively high effluent COD, iron and suspended solids concentrations, and low UV transmittance. This was confirmed herein. For UV, the inactivation curve for fecal coliforms showed the typical two-stage shape, with the target of 1000 CFU/100 mL (to account for photoreactivation) occurring in the asymptote zone at fluences >20 mJ/cm(2). In contrast, inactivation curves for MS-2 and Clostridium perfringens were linear. Clostridium perfringens was the most resistant organism. For ozone, inactivation was already observed before any residuals could be measured. The transferred ozone doses to reach target fecal coliform levels ( approximately 2-log reduction) were 30-50 mg/L. MS-2 was less resistant, but Clostridium perfringens was more resistant than fecal coliforms. The different behaviour of the four indicator organisms studied, depending on the disinfectant, suggests that a single indicator organism might not be appropriate. The required dose of any of the disinfectants is unlikely to be economically viable, and upstream changes to the plant will be needed.     

Some effects of pulp and paper wastewater on microbiological water quality of a river

A study was conducted to detect the source of fecal indicator bacteria and measure the resulting impairment of water quality of the Sturgeon River at Sturgeon Falls, Ontario, Canada. The bacteriological water quality of the river above the dam at Sturgeon Falls was fairly good, while below the dam the water quality was degraded. The principal input of fecal indicator bacteria was traced to the paper mill at Sturgeon Falls. The impairment of water quality was detected all the way to recreational areas near the mouth of the river. The impaired stretch of the river had diminished aesthetic appeal. Other sources of indicator bacteria could be masked by the large numbers of similar bacteria discharged from the paper mill. The principal fecal coliform was Klebsiella pneumoniae. In pulp mill wastes the fecal origin of this bacterium can be disputed, and so its presence in large numbers interfered with the interpretation of fecal coliform results. K. pneumoniae is also an opportunistic pathogen which causes infections in humans. Thermotolerant oxidase positive bacteria which were isolated from the paper mill wastes, register as false positives in the fecal coliform test causing problems in interpretation. Pseudomonas aeruginosa was detected in the pulp mill wastewater, and at a level of 82 PA 100 ml⁻¹ in recreational areas of the river. This was considered to be of some hazard to users of the water. In addition, Escherichia coli was detected in the mill wastewater, at about 200 EC 100 ml⁻¹, and this probably indicated some measure of fecal pollution though likely of animal origin. Finally the nutrient rich wastewater led to an approx. 50-fold increase in density of aerobic heterotrophic bacteria in the river water, as well as the production of a slimy filamentous growth on surfaces, stones and wooden pilings, in the river. The principal organism in this slime was the fungus Leptomitus.     

Bacterial population changes in hospital effluent treatment plant in central India

Hospital effluent with its high content of multidrug resistant (MDR) enterobacteria and the presence of enteric pathogens could pose a grave problem for the community. It was planned at our tertiary care hospital in central India to study the population changes at various steps of effluent treatment plant (ETP) like collection, aeration, clarification, liquid sludge, dried sludge, high-pressure filter and treated wastewater. The study included viable bacterial counts, coliform counts, staphylococcal, enterococcal, Pseudomonas and multiple drug resistant (MDR) gram negative bacterial counts in the different stages of ETP. In order to study the distribution of bacteria as free floating in liquid and adherent to suspended particles, enumeration of the bacteria in the filterate and the sediment was also carried out. The effluent input showed 55% of the 8.6 x 10(6)/ml bacteria as coliforms and E. coli which was a typical of fecal flora. The prevalence of MDR coliforms was 0.26%. The substantial reduction (> 3log) was seen for the effluent coming from the clarifier. The bulk of the bacteria in the hospital effluent remains firmly adhered to solid particles; aeration and clarification removes bulk of the bacteria by physical processes like flocculation. The treated liquid effluent still contains sizeable loads of MDR bacteria and inactivation by procedure such as chlorination is required. The bacteria get concentrated in sludge and a greater concentration of chlorine is required for decontamination.     

Detection of the human specific Bacteroides genetic marker provides evidence of widespread sewage contamination of stormwater in the urban environment

Human sewage contamination of surface waters is a major human health concern. We found urban stormwater systems that collect and convey runoff from impervious surfaces act as a conduit for sewage originating from breeches in sanitary sewer infrastructure. A total of 828 samples at 45 stormwater outfalls were collected over a four-year period and assessed by culture based methods, PCR, and quantitative PCR (qPCR) to test for traditional and alternative indicators of fecal pollution. All outfalls had the HF183 (human) Bacteroides genetic marker detected in at least one sample, suggesting sewage contamination is nearly ubiquitous in the urban environment. However, most outfalls were intermittently positive, ranging from detection in 11%-100% of the samples. Positive results did not correlate with seasonality, rainfall amounts, or days since previous rainfall. Approximately two-thirds of the outfalls had high (>5000 copy number, i.e. CN, per 100 ml) or moderate levels (1000-5000 CN per 100 ml) of the human Bacteroides genetic marker. Escherichia coli (E. coli) and enterococci levels did not correlate to human Bacteroides. A total of 66% of all outfall samples had standard fecal indicator levels above 10,000 CFU per 100 ml. A tiered assessment using this benchmark to identify high priority sites would have failed to flag 35% of the samples that had evidence of sewage contamination. In addition, high fecal indicators would have flagged 33% of samples as priority that had low or no evidence of sewage. Enteric virus levels in one outfall with high levels of the human Bacteroides genetic marker were similar to untreated wastewater, which illustrates stormwater can serve as a pathway for pathogen contamination. The major source of fecal pollution at four of five river sites that receive stormwater discharge appeared to be from sewage sources rather than non-human sources based on the ratios of human Bacteroides to total Bacteroides spp. This study shows the feasibility and benefits of employing molecular methods to test for alternative indicators of fecal pollution to identify sewage sources and potential health risks and for prioritization of remediation efforts.     

BOD5 estimation for pulp and paper mill effluent using UV absorbance

A novel method based on UV absorbance, is presented for estimating the BOD5 in pulp and paper mill effluent. This method could eventually be incorporated into an on-line sensor for BOD5 that is suitable for process control applications. Two streams, the reactor entrance and the final effluent, from two different mills were studied. One mill employed the Kraft pulping process, while the second mill was a thermo-mechanical one. The absorbance over the range 200-350 nm showed significant differences between the two mills. Because the two mills use very distinct processes, separate correlations were used to relate the absorbance to the BOD5 for both the mills. Results indicate that prediction of reactor entrance BOD5 was reasonable, whereas prediction of final effluent BOD5 was inaccurate, for both mills. Also studied was the effect of aeration on BOD5 results obtained at low BOD5 values for the Kraft mill.     

Fecal bacteria and sex hormones in soil and runoff from cropped watersheds amended with poultry litter

The application of poultry litter to agricultural fields can provide plant nutrients for crops and forage production, but fecal bacteria and the sex hormones estradiol and testosterone are components of litter that can be detrimental to the environment. Our objective was to determine if applications of poultry litter to small watersheds would contribute to the load of fecal bacteria and sex hormones to soil and runoff. We, therefore, investigated the fate and transport of fecal bacteria, estradiol and testosterone from surface applied poultry litter to four small cropped watersheds. Poultry litter was applied to meet the nitrogen requirements of pearl millet (Pennisetum glaucum L.) in 2000 and grain sorghum [Sorghum bicolor (L.) Moench] in 2001. Neither Salmonella nor Campylobacter were detected in the litter but the fecal indicator bacteria were. The average load of total coliforms, Escherichia coli, and fecal enterococci applied with the litter was 12.2, 11.9, and 12.7 log(10) cells ha(-1), respectively. The average load of estradiol and testosterone was 3.1 and 0.09 mg ha(-1), respectively. Runoff events first occurred 7 months after the first litter application in 2000, and 3 weeks after the second application in 2001. Only for the 25 July 2001 runoff event 3 weeks after the second litter application were the concentrations of total coliforms, E. coli, and fecal enterococci in runoff greater than background concentrations which were on average 5.2, 2.9, and 1.1 log(10) MPN 100 ml(-1), respectively. Average background levels of total coliforms, fecal enterococci, and E. coli in surface soil were 8.2, 7.9, and 3.5 log(10) cells kg(-1) soil. At the rate of litter application the concentrations of estradiol and testosterone in the litter did not appear to impact the background levels in the soil and runoff. Because concentrations of sex hormones in litter from other broiler operations are known to be greater than in the litter we applied, further study on the connection between concentrations of sex hormones in poultry litter and operational practices is recommended.     

Microbiological effects of wastewater effluent discharge into coastal waters of Puerto Rico

Microbiological effects of non-disinfected, combined chemical and domestic wastewater effluent discharge into coastal waters north of Barceloneta. Puerto Rico were investigated by membrane filter enumeration of fecal indicator bacteria and by enrichment isolation of specific pathogens. A wastewater plume was detected and delineated around the sewage outfall, located 800 m offshore and 30 m below the ocean surface. Fecal coliform and fecal streptococcus counts within the plume were significantly (F test) larger than comparable counts outside the plume. The shape and location of the plume was compatible with current patterns and prevailing winds and extended as far as 3.7 km west of the outfall, at which point the fecal coliform count was 146 100 ml⁻¹. The average non-plume fecal coliform concentration was 6.7 100 ml⁻¹. Specific pathogens isolated included Salmonella and Vibrio spp, including V. cholerae non-01 serovar isolated from a nearby river also impacted by the sewage plant.     

Regrowth of coliforms and fecal coliforms in chlorinated wastewater effluent

Observations made both in the field in chlorinated effluent, and in laboratory experiments show that coliforms and fecal coliforms are capable of regrowth in chlorinated wastewater. Under field conditions regrowth of coliforms in chlorinated effluent held in a storage reservoir for about 3 days appeared inversely correlated to: (1) The residual chlorine in the storage reservoir and (2) The number of coliforms surviving chlorination. In the laboratory experiments regrowth occurred after initial doses as high as 11 ppm total chlorine even when there was no chemical inactivation of the chlorine. Fecal coliforms did not generally show regrowth to the same extent as coliforms. Regrowth occurred even when coliforms were not detectible in 10-ml of samples after chlorination.Since coliforms and fecal coliforms are capable of regrowth in chlorinated sewage effluent and admixtures of it, the sanitary significance of the number of coliforms after storage or in receiving bodies of water is difficult to interpret. Thus standards might be based on the number of coliforms, or fecal coliforms detected in effluents immediately after chlorination. However, this would not be justified if in addition to coliforms, pathogenic bacteria can regrow in chlorinated effluents.     

North and south american studies on the potential of coliphage as a water quality indicator

Studies were undertaken to assess the potential of coliphages to be used universally as water quality indicators and more specifically as health hazard indicators. Data were obtained from three water bodies, a northern Canadian River, inshore water samples from Lake Ontario and from marine beaches in Brazil. Data from this two continent, three water body study indicated (a) that within location fecal coliform and coliphages are positively correlated, (b) coliphage values can be predicted by using fecal coliform MPN, fecal streptococci MF and E. coli MF data and (c) a water quality guideline of 20 coliphage/100 ml for recreational fresh waters is proposed.     

Assessment of sources of human pathogens and fecal contamination in a Florida freshwater lake

We investigated the potential for a variety of environmental reservoirs to harbor or contribute fecal indicator bacteria (FIB), DNA markers of human fecal contamination, and human pathogens to a freshwater lake. We hypothesized that submerged aquatic vegetation (SAV), sediments, and stormwater act as reservoirs and/or provide inputs of FIB and human pathogens to this inland water. Analysis included microbial source tracking (MST) markers of sewage contamination (Enterococcus faecium esp gene, human-associated Bacteroides HF183, and human polyomaviruses), pathogens (Salmonella, Cryptosporidium, Giardia, and enteric viruses), and FIB (fecal coliforms, Escherichia coli, and enterococci). Bayesian analysis was used to assess relationships among microbial and physicochemical variables. FIB in the water were correlated with concentrations in SAV and sediment. Furthermore, the correlation of antecedent rainfall and major rain events with FIB concentrations and detection of human markers and pathogens points toward multiple reservoirs for microbial contaminants in this system. Although pathogens and human-source markers were detected in 55% and 21% of samples, respectively, markers rarely coincided with pathogen detection. Bayesian analysis revealed that low concentrations (<45 CFU × 100 ml⁻¹) of fecal coliforms were associated with 93% probability that pathogens would not be detected; furthermore the Bayes net model showed associations between elevated temperature and rainfall with fecal coliform and enterococci concentrations, but not E. coli. These data indicate that many under-studied matrices (e.g. SAV, sediment, stormwater) are important reservoirs for FIB and potentially human pathogens and demonstrate the usefulness of Bayes net analysis for water quality assessment.     

Microbial pollution indicators in Brazilian tropical and subtropical marine surface waters

The specificity of indicators that depend on elevated incubation temperature as a selective factor for their enumeration was questioned because of the possibility of interference from autochthonous microorganisms adapted to high ambient temperatures. Lactose-fermenting cultures isolated from fecal coliform tests of tropical marine surface waters were identified as consisting of about 70% Escherichia coli, and most of the remaining cultures being Klebsiella, Enterobacter or Citrobacter species. This confirmed the taxonomic specificity of fecal coliform tests for these waters. Fecal and total coliforms, fecal streptococcus, heterotrophic bacteria and yeast counts had correlations of above 99% confidence levels with most other microbial and chemical parameters studied. Waters with fecal coliform counts above 1000 per 100 ml had increased incidence of presumptive pathogenic yeasts, Pseudomonas aeruginosa and Salmonella. Our data support the use of coliforms or fecal streptococci as indicators of recent fecal pollution in tropical marine waters and yeast or heterotrophic bacteria counts as complementary indicator methods for these waters.     

Fecal indicator bacteria are abundant in wet sand at freshwater beaches

Potential fecal contamination of sand in the wave-washed zone of public bathing beaches is overlooked in beach monitoring programs. Activity in this zone can bring pathogens to the sand surface or into the water, presenting a health risk to sensitive populations. On a unit weight basis (colony forming units per 100g), the mean summer abundance of the fecal indicator bacteria enterococci and Escherichia coli was 3-38 times higher in the top 20 cm of wet-sand cores than in the water column at six freshwater bathing beaches. E. coli were 4 times more abundant than enterococci in water but counts were similar in the sand. A correlation (r=0.60) existed between E. coli counts in the water and in the top 5 cm of sand only, whereas no relationship existed between enterococci abundance in water and sand. In general, enterococci were most numerous in the 5-10 cm sand stratum and E. coli in the 0-5 cm stratum. These preliminary data show that wet freshwater beach sand is a reservoir of fecal indicator bacteria. Enteric pathogens may also be present in beach sand.     

Fate of indicator microorganisms, Giardia and Cryptosporidium in subsurface flow constructed wetlands

Limited information is available on the ability of subsurface flow wetlands to remove enteric pathogens. Two multi-species wetlands, one receiving secondary sewage effluent and the other potable (disinfected) groundwater were studied from February 1995 to August 1996, at the Pima County Constructed Ecosystems Research Facility in Tucson, Arizona. Each wetland had a retention time of approximately 4 days. The objectives of this study were (1) to evaluate the ability of multi-species subsurface wetlands to physically remove Giardia cysts; Cryptosporidium oocysts, total and fecal coliforms, and coliphages; and (2) to determine the likely impact of local wildlife on the occurrence of these indicators and pathogens. In the wetland receiving secondary sewage effluent, total coliforms were reduced by an average of 98.8% and fecal coliforms by 98.2%. Coliphage were reduced by an average of 95.2%. Both Giardia cysts and Cryptosporidium oocysts were reduced by an average of 87.8 and 64.2%, respectively. In the wetland receiving disinfected groundwater, an average of 1.3 x 10(2) total coliforms/100 mL and 22.3 fecal coliforms/100 mL were most likely contributed by both flora and fauna. No parasites or coliphages were detected.     

In situ persistence of indicator bacteria in streams polluted with acid mine drainage

Persistence of indicator organisms (total coliforms, fecal coliforms, and fecal streptococci) associated with natural samples of raw sewage was studied following in situ exposure to five aquatic environments. Three of these streams contained significant amounts of acid mine water (AMW) while the other two were relatively uncontaminated. Indicator organisms were rapidly killed upon exposure to the acid mine systems, whereas little reduction in numbers was observed in the uncontaminated streams. Seasonal changes affected survival of indicators as reflected by prolonged persistence at colder in situ water temperatures. The fecal coliform group was most susceptible to the AMW stress, while the fecal streptococci were most persistent. An enrichment technique resulted in substantially enhanced recovery of certain species of sublethally injured survivors of acid stress. Enrichment was particularly beneficial for recovery of AMW-injured fecal coliforms. Relatively little improvement in recovery of fecal streptococci was afforded by the enrichment technique.     

Fate of physical,chemical, and microbial contaminants in domestic wastewater following treatment by small constructed wetlands

In order to evaluate the efficacy of constructed wetlands for treatment of domestic wastewater for small communities located in rural areas, small-scale wetland mesocosms (400 L each) containing two treatment designs (a mixture of Typha, Scirpus, and Juncus species; control without vegetation) were planted into two depths (45 or 60 cm) with pea gravel. Each mesocosm received 19 L/day of primary-treated domestic sewage. Mesocosms were monitored (inflow and outflow samples) on a monthly basis over a 2-year period for pH, total suspended solids (TSS), 5-day biochemical oxygen demand (BOD(5)), total Kjeldahl nitrogen (TKN), dissolved oxygen (DO), and conductivity. Microbiological analyses included enumeration of fecal coliforms, enterococci, Salmonella, Shigella, Yersinia, and coliphage. Significant differences between influent and effluent water quality for the vegetated wetlands (p<0.05) were observed in TSS, BOD(5), and TKN. Increased DO and reduction in fecal coliform, enterococcus, Salmonella, Shigella, Yersinia, and coliphage populations also were observed in vegetated wetlands. Greatest microbial reductions were observed in the planted mesocosms compared to those lacking vegetation. Despite marked reduction of several contaminants, wetland-treated effluents did not consistently meet final discharge limits for receiving bodies of water. Removal efficiencies for bacteria and several chemical parameters were more apparent during the initial year compared to the second year of operation, suggesting concern for long-term efficiency and stability of such wetlands.